ABSTRACT Through studies proposed here, we will develop a strategy of ?negative immunization? to achieve antigen-specific tolerance for the prevention and immunotherapy of atopic dermatitis (AD), a highly common chronic inflammatory skin disease that presents a considerable socioeconomic burden. We intend to manipulate neuroimmune regulatory networks in the skin to prevent and treat pathologic innate and adaptive immune responses accounting for the acute and chronic phases of AD. The novel ?negative immunization? approach has been engineered to prevent the priming of nave T and B cells and to eradicate existing allergen specific T- and B- cell effector responses. To accomplish this, we will administrate allergen/antigen together with antagonists of the neurokinin-1 receptor (NK1R) to prevent receptor binding by its inflammatory agonists, an interaction that is critical for the initiation of innate and adaptive effector and memory immune functions. The rationale for this approach is that eliminating/decreasing inflammation at the time of skin allergen/Ag encounter will maintain a quiescent a microenvironment for antigen presenting cells (APCs). This will result in the generation of anti- inflammatory APCs capable of presenting the encountered antigen to T cells in a tolerogenic fashion. This strategy will enable, for the first time, an antigen specific strategy for the prevention and treatment of AD. We hypothesize that: ?Promoting an anti-inflammatory cutaneous microenvironment by efficient co- delivery of allergen/Ag and NK1R antagonists will generate anti-inflammatory APCs to tolerize Ag specific T cells and mitigate/abrogate pre-existing memory B and T-cell responses that cause AD?. The negative immunization approach will be administered, utilizing novel and enabling microneedle arrays (MNAs) developed in our laboratories. These MNAs integrate and release biologically active agents, with highly replicable dosage control and have been formulated to achieve simultaneous delivery of allergen / Ag and neuroimmunomodulatory small molecules to a specific skin stratus. Combining this innovative immunoregulatory approach with our novel MNA delivery technology, will enable us to engineer the cutaneous microenvironment ?in vivo? for the prevention and therapy of AD. Importantly, our experiments include translational studies focusing on human skin that are specifically designed to enable rapid translation of this strategy to clinical trials.